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Construction method and application of recombinant lactobacillus acidophilus for expressing glucagon-like peptide-2

A technology of glucagon-like peptide and Lactobacillus acidophilus, applied in the field of animal husbandry and veterinary medicine, can solve problems such as ineffective repair and intestinal damage, and achieve the effects of repairing intestinal loss, promoting animal growth and improving intestinal environment.

Active Publication Date: 2015-03-04
SHENZHEN JINXINNONG FEED
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Problems solved by technology

[0008] The purpose of the present invention is to provide a construction method and application of recombinant Lactobacillus acidophilus, and to develop a method that can promote Recombinant Lactobacillus acidophilus for intestinal mucosal injury repair in piglets

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  • Construction method and application of recombinant lactobacillus acidophilus for expressing glucagon-like peptide-2
  • Construction method and application of recombinant lactobacillus acidophilus for expressing glucagon-like peptide-2
  • Construction method and application of recombinant lactobacillus acidophilus for expressing glucagon-like peptide-2

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[0022] See attached Figure 1-Figure 4 The recombinant Lactobacillus acidophilus expressing porcine glucagon-like peptide-2 described in the present invention is constructed by the following method: according to the published gene sequence of porcine glucagon-like peptide-2 (glp2) and the expression vector plasmid Features: design primers containing specific enzyme cutting sites; use porcine small intestinal mucosa as material, extract mRNA, reverse transcribe cDNA as template for PCR amplification, obtain glp2 gene fragment, and connect it to the expression vector plasmid pMG36e , the recombinant plasmid pMG36e-glp2 was obtained, and the recombinant plasmid was electrotransformed into Lactobacillus acidophilus competent cells by an electroporation method for expression. Animals were fed with the constructed recombinant porcine glucagon-like peptide-2 Lactobacillus acidophilus, and the effect of preventing and treating diarrhea was observed. Required experimental material of ...

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Abstract

The invention discloses a construction method and application of recombinant lactobacillus acidophilus for expressing glucagon-like peptide-2. The construction method comprises the following steps: designing a primer containing specific restriction enzyme digestion site according to gene sequence and expression vector plasmid characteristic of disclosed glucagon-like peptide-2; by using small intestinal mucosa as a material, carrying out RT-PCR (reverse transcription-polymerase chain reaction) to obtain fragment containing glucagon-like peptide-2 gene; connecting the fragment with lactobacillus expression vector plasmid Pmg36e to obtain recombinant plasmid Pmg36e-glp2; transferring the recombinant plasmid in actobacillus acidophilus through an electrotransformation method to obtain recombinant lactobacillus acidophilus for expressing glucagon-like peptide-2; expressing under the induction of nisin; and analyzing and proving that the gene is correctly expressed through SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and Western blot. The recombinant lactobacillus acidophilus is used for preventing and treating diarrhea, improving the intestinal health of pig and improving the growth performance.

Description

technical field [0001] The invention belongs to the technical field of animal husbandry and veterinary medicine, and in particular relates to a construction method of recombinant Lactobacillus acidophilus expressing porcine glucagon-like peptide-2 and its application in promoting intestinal health of pigs. Background technique [0002] At present, the pig industry has become an important industry related to the national economy and the people's livelihood. In pig production, piglets are the stage with the fastest growth and development, the highest feed utilization rate, and the greatest development potential. The quality of piglet feeding directly affects the production performance of pigs throughout the feeding period. However, the piglet stage is also the most vulnerable stage of viability and physiological function, and the mortality rate of piglets usually accounts for 70% of the mortality rate in the whole feeding period. The low survival rate and slow growth of pigle...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/74C12N1/21A23K1/18A23K1/16C12R1/23
Inventor 侯永清吴涛窦茂鑫赵迪丁斌鹰
Owner SHENZHEN JINXINNONG FEED
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